LOCUS       sequence-403941-        4697 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Ash-tagged LC3B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4697)
  AUTHORS   Ishimura R, El-Gowily AH, Noshiro D, Komatsu-Hirota S, Ono Y, Shindo
            M, Hatta T, Abe M, Uemura T, Lee-Okada HC, Mohamed TM, Yokomizo T, 
            Ueno T, Sakimura K, Natsume T, Sorimachi H, Inada T, Waguri S, Noda 
            NN, Komatsu M
  TITLE     The UFM1 system regulates ER-phagy through the ufmylation of CYB5R3.
  JOURNAL   Nat Commun. 2022 Dec 21;13(1):7857. doi: 10.1038/s41467-022-35501-0.
  PUBMED    36543799
REFERENCE   2  (bases 1 to 4697)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2022-12-21"; volume: "13(1):7857. doi"; pages: " 
            10.1038/s41467-022-35501-0"
COMMENT     Sequence Label: pAsh-MCL-LC3B
FEATURES             Location/Qualifiers
     source          1..4697
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    881..1002
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(918..937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     972..991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1009..1464)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1146..1167)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1358..1377
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1491..1595
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1597..1954
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1597..1788
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1620..1640)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1805..1940
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1988..2782
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1988..2782
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2042..2061)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2652..2671
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2970..2989)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3014..3061
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3390..3978
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3879..3898
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4153..4456
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4457..4660
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4611..4631
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      join(4692..4697,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggcgtcgc tcaccgtgaa ggcctacctt ctgggcaagg aggacgcggc gcgcgagatt
       61 cgccgcttca gcttctgttg cagccccgag cctgaggcgg aagccgaggc tgcggcgggt
      121 ccgggaccct gcgagcggct gctgagccgg gtggccgccc tgttccccgc gctgcggcct
      181 ggcggcttcc aggcgcacta ccgcgatgag gacggggact tggttgcctt ttccagtgac
      241 gaggaattga caatggccat gtcctacgtg aaggatgaca tcttccgaat ctacattaaa
      301 gagaaaaccg gtaattccgc tgacggcggc ggaggatcgg gtggtagtgg tggttcagga
      361 ggaggatcga cccaaggagg atccatgccg tcggagaaga ccttcaagca gcgccgcacc
      421 ttcgaacaaa gagtagaaga tgtccgactt attcgagagc agcatccaac caaaatcccg
      481 gtgataatag aacgatacaa gggtgagaag cagcttcctg ttctggataa aacaaagttc
      541 cttgtacctg accatgtcaa catgagtgag ctcatcaaga taattagaag gcgcttacag
      601 ctcaatgcta atcaggcctt cttcctgttg gtgaacggac acagcatggt cagcgtctcc
      661 acaccaatct cagaggtgta tgagagtgag aaagatgaag atggattcct gtacatggtc
      721 tacgcctccc aggagacgtt cgggtaaaag cttgaataag cggccgcgac tctagatcat
      781 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
      841 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
      901 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
      961 gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa attgtaagcg
     1021 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     1081 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     1141 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     1201 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     1261 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     1321 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     1381 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     1441 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     1501 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     1561 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     1621 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     1681 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     1741 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     1801 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     1861 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     1921 aggaggcttt tttggaggcc taggcttttg caaagatcat caagagacag gatgaggatc
     1981 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     2041 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     2101 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     2161 tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     2221 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     2281 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     2341 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     2401 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     2461 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat
     2521 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     2581 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     2641 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     2701 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     2761 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     2821 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     2881 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     2941 ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat accggaagga
     3001 acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt
     3061 cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat
     3121 tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa
     3181 ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca
     3241 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     3301 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3361 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3421 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3481 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     3541 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3601 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3661 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3721 tgcacacagc ccagtttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3781 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3841 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3901 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3961 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4021 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     4081 accgccatgc attagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     4141 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     4201 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     4261 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     4321 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     4381 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     4441 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     4501 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4561 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     4621 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat
     4681 ccgctagcat tgccacc
//
